Study Of Heme Oxygenase-1 On Human Mesenchymal Stem Cells.

Objective:To estabish methods of culture and identidy of MSCs and purification of peripheral blood T lymphocytes in vitro,and study the presence of heme oxygenase-1 (HO-1) of mesenchymal stem cells and the effect in mesenchymal stem cells apoptosis. To investigate the influence of immune cells and immune regulator act on human mesenchymal stem cells when heme oxygenase-1 absence.As well as to investigate the role for HO-1 in the immunosuppressive effect of mesenchymal stem cells.Methods:Human mesenchymal stem cells were cultured in vitro.MSCs were isolated from human bone marrow and cutuled.The purity of them were detected by flow cytometry.T-lymphocytes were harvested by using nylon column.The method of RT-PCR were used to detect the expression of HO-1mRNA.Morphology observation and flow cytometry Annexin V were used to detect apoptosis and to dertermine the content of DNA.RT-PCR method was used to detect HO-1 mRNA expression and flow cytometry was used to analyze cell apoptosis with the stimulation of IFN-γand T cells activated by PHA.MTT was used to analyze T lymphocyte proliferation and the celluar proliferation inhibition rate was calculated.Results:Heme oxygenase-1 mRNA was expressed in mesenchymal stem cells of normal bone marrow,apoptosis was promoted by adding Znpp-Ⅸ,the peak of apoptosis was clear on the distribution figerature of DNA.Heme oxygenase-1 mRNA was descended with the stimulation of IFN-γand activated T cells.Adding IFN-γand Znpp-Ⅸ, the mesenchymal stem cells apoptosis increased.While both of them added,the MSC apoptosis was more obvious.MSC could inhibite proliferation of the allogenetic T-inhibition lymphocytes induced by PHA.When adding Znpp-Ⅸ,the immunosuppressive effect of T-lymphocytes affected by MSC would be decreased. Conclusion:HO-1 played an anti-apoptosis role in human mesenchymal stem cells. HO-1 played an important role in the immunosuppressive effect of human mesenchymal stem cells.Immune cells and immune regulator made the expression of HO-1mRNA in mesenchymal stem cells descending and the experimental results provided a new theoretical basis to study the pathogenesis of aplastic anemia.